Waters MassPREP Phosphopeptide Enrichment Kit - Manual

[ Care and Use ManUal ]
I. Int roductIon
The MassPREP ™ Phosphopeptide Enrichment Kit is a tool for selective
enrichment of phosphopeptides from complex proteomic samples. This
kit contains a micro-scale solid-phase extraction (SPE) plate packed with
a unique affinity sorbent. This affinity SPE does not require pre-loading
the sorbent with metal chelation. Also, superior selectivity is achieved
compared to other existing affinity chromatographic methods.
The selectivity of this sorbent towards phosphopeptides is further
improved by adding a unique chemical to the sample before loading
onto the SPE. This chemical (under the trade name, Enhancer) improves
the overall selectivity by displacing the non-phosphorylated peptides
during the sample loading step.
The MassPREP ™ Phosphopeptide Enrichment Kit includes the 96-well
micro elution (µElution) plate, enhancer, and a test sample for method
validation. The test sample is a mixture of four synthetic phosphopeptides
mixed with yeast enolase tryptic peptides in 1:1 molar ratio. A general
guideline for kit operation and specific protocols for phosphopeptide
extraction from complex protein digests are provided. The 96-well
micro elution (µElution) platform conveniently uses a vacuum manifold,
for reproducibility and high throughput of enrichment experiments.
contEntS
I. INTRODUCTION
II. MATERIALS INCLUDED IN THE KIT
III. PHOSPHOPEPTIDE AFFINITY ENRICHMENT METHOD
IV. PHOSPHOPEPTIDE ENRICHMENT KIT USAGE GUIDELINES
Part A. How to Use the MassPREP ™ Phosphopeptide Enrichment
μElution Plate
Part B. How to Use the MassPREP ™ Enolase/Phosphopeptide
Standard as a Test Sample
Part C. How to Apply Enhancer for Highly Complex Samples
Part D. Sample Preparation for Mass Spectrometry Analysis
V. ORDERING INFORMATION
VI. STORAGE AND DISPOSAL OF USED PLATES
MassPrEP ™ PhoSPhoPEPtIdE EnrIchMEnt KIt